The Tissue Procurement and Cell Culture Core
Director: Scott H. Randell, PhD
•Lung Cell Isolation and Culture, Including Primary Airway and Alveolar Epithelium, Microvascular Endothelial Cells and Fibroblasts
•Specialized Media Formulations
•In vivo and in vitro Airway Surface Materials
•Genetic Manipulation of Lung Cell Cultures
•Creation and Characterization of Novel Cell Lines
The Molecular Biology Core
Director: Wanda K O’Neal, PhD
•The Core serves as a centralized resource for cloning and sequencing of genes.
•Isolation, storage, and analysis of high quality RNA.
•Generation of transgenic mouse models.
•Target validation– development of methods for si/shRNA, CRISPR knockdown in airway cell models.
•Provide training in molecular biology.
The Histology Core
•Light microscopy services include: prosection, tissue fixation, processing, embedding, sectioning, staining and cover slipping of paraffin, frozen and soft plastic tissue sections.
•The histology core also provides expertise in all aspects of transmission and scanning electron microscopy. Access to a high pressure freezer and automated freeze substitution is available.
•A highlight of the core facility is our specialized procedure for histological evaluation of airway cells grown on porous membrane supports.
CFTR Functional Analysis Core and Cystic Fibrosis Molecular/Functional Measurement Core
Director: Martina Gentzsch, PhD
•In vitro short-circuit current measurements to assess ion channel properties in primary and conditionally reprogrammed bronchial and nasal epithelial cells from patients (CF, COPD etc.) and controls.
•Ex vivo measurements of ion transport function in rectal biopsies from patients and controls.
•In vivo NPD, salivary secretion, and mucociliary clearance measurements in mouse models.
•Analysis of CFTR protein expression and biochemical correction of mutant CFTR.
•Assessment of efficacy of functional rescue of mutant CFTR by pharmacological agents.
•Advanced and Standard Microscopy
•Live Cell Imaging
•Color Brightfield Imaging (high resolution)
•Stereo Fluorescence Micro/Macroscope
•Laser Micro-Dissection System
•Image Processing and Analysis
The Animal Models Core
Director: Wanda K O’Neal, PhD
Co-Director: Dr. Alessandra Livraghi-Butrico, PhD
•The Mouse Models Lab has developed and maintains active colonies of various mouse models relevant for basic and translational pulmonary research, i.e., βENaC (Scnn1b) transgenic, ΔF508 Cftr mice, secreted and tethered Mucin-deficient mice, and relative intercrosses. Models of acute lung injury, sepsis, pneumonia/infection, cigarette smoke exposure are also available.
•Phenotyping: bronchoalveolar lavage (neonates and adults), differential cell counts and inflammatory markers, histopathology and morphometry, secreted mucins agarose western blots, airway mucus burden, microbiology, airway mucus clearance measurements.
•Administration of pharmacologic treatments via intra-tracheal instillation, nasal aspiration, gavage, subcute or intraperitoneal injection, in neonatal or adult mice.
Center for Airways Disease
Director: Claire Doerschuk, MD
•Recruitment of leukocytes during pneumonia and other lung injuries.
•Endothelial cell biology in normal lungs and during inflammatory and innate immune processes.
•Cigarette-smoke induced lung disease.
•Effector functions of leukocytes.
•The function of the small GTPase Rac2 expressed in endothelial cells which regulates both normal fluid flux and edema formation during injury.
Mucociliary Transport (MCT) and Mucus Clearance
–Airway Surface Liquid (ASL) depth, mucociliary transport, ciliary beat frequency
–Penetration kinetics of drugs through mucus
–Mucus rheology, biophysics and clearance
–Tools for mucolytic drug screening
•Mouse in-vivo MCT
•MCT in human volunteers